Fig. 2 –

Induction of Armet expression by chemical ER stress inducers in cells and by ischemia in rat brain. A. Armet is upregulated by different ER stress inducers in different cell lines. U2OS, HEK293 and SHSY-5Y were treated with tunicamycin (Tun, 2.5 μg/ml), thapsigargin (Tha, 500 nM), or proteasome inhibitor lactacystin (Lac, 10 μM) for 7 h. B and C. Armet is upregulated by ER stress in a time-dependent manner. D. Armet but not Armet-L1 is upregulated by tunicamycin. U2OS cells were treated with Tunicamycin for 0, 1, 2, 5 and 7 h. Total RNA was extracted and RT-PCR was performed. E. Anti-Armet antibody is specific for Armet in immunostaining. U2OS cells were transfected with pCIneo-Armet-FLAG. Twenty-four hours after transfection cells were fixed and stained with rabbit polyclonal anti-Armet and mouse monoclonal anti-FLAG antibodies in conjunction with Alexa Fluor-488 (green) and Alexa fluor-594 (red). F and G. Induction of Armet by experimental cerebral ischemia. Armet was revealed by immunohistochemical staining by ABC method of ischemic cerebral cortex (G) or contralateral side (F, negative control). Nuclei were counterstained by hematoxylin.